Chemical signals govern a large part of insect behavior and
physiology. Within a species chemical communication is mediated by
pheromones. Insect pheromones are described briefly, with particular
emphasis on sex attractants. Identification of pheromones is presented
using the example of an unusual, caffeine-like alkaloid,
1,3-dimethyl-2,4-(1H,3H)-quinazo-linedione, the sex pheromone of the
pale-brown chafer, Phyllophertha diversa. Biosynthetic pathways are
described for pheromones from the best studied insect orders
(Lepidoptera, Coleoptera, Diptera and Dictyoptera). The methodology for
establishing the biosynthetic routes is presented using unsaturated g
-lactones, the sex pheromones of a number of scarab beetles. Evidence
is shown that these semiochemicals are biosynthesized from fatty acids
by desaturation, hydroxylation, chain shortening and cyclization
reactions. The hydroxylation reaction is remarkably stereospecific in
two species: the Osaka beetle (Anomala osakana) and the Japanese beetle
(Popillia japonica). Molecular mechanisms of pheromone perception are
presented, with the emphasis on the perireceptor events, taking place
in the sensillar lymph. Pheromone binding proteins (PBPs), transporting
the pheromones from the environment to the receptor neurons, are
described in detail. Purification, cloning, and ligand binding
specificity of two new classes of these proteins identified in scarab
beetles are described. Preparation of the recombinant PBP from the
silkworm moth, Bombyx mori, expressed in Escherichia coli periplasm, is
presented. Structural changes induced by pH change in this protein and
the odorant binding protein from P. diversa, and a similar
conformational transition in B. mori PBP in the pre-sence of model
phospholipid membranes, are described. A novel mechanism of ligand
transport through the sensillar lymph and its release upon encountering
the surface of olfactory neurons is proposed based on these structural
studies. Enzymatic inactivation of pheromones by sensillar enzymes is
described. Degradation of g -lactones in several species of scarab
beetles is analyzed and antennal esterases metabolizing these compounds
are characterized. Substantial substrate specificity for the natural
stereoisomer is demonstrated in two cases (Anomala octiescostata and P.
japonica). Degradation pathway of
1,3-dimethyl-2,4-(1H,3H)-quinazolinedione is also analyzed. Evidence is
presented that this molecule is metabolized by a specific, olfactory
cytochrome P450 by N-de-methylation and hydroxylation of the aromatic
ring. Potential applications for results of molecular studies of insect
chemical communication are discussed.
Spis treści:
Table of contents
List of abbreviations
1. Introduction
2. Insect pheromones
2.1. Structural diversity and species specificity
2.2. Identification of pheromones
3. Pheromone biosynthesis
3.1. Anatomical location of pheromone biosynthesis
3.2. Pheromone biosynthesis in model insect species
3.3. Biosynthesis of y-lactones in scarab beetles
3.4. Hormonal regulation of pheromone biosynthesis
4. Pheromone perception in the olfactory sensillum
4.1. Structure of the olfactory sensillum
4.2. Biochemical events in the olfactory sensillum
4.3. Pheromone binding proteins
4.3.1. Pheromone binding proteins in scarab beetles, P, japonica and A. osakana - the problem of ligand binding specificity
4.3.2. Odorant binding proteins in the pale brown chafer, P. diuersa - the problem of heterogeneity
4.3.3. The function of structural changes in pheromone binding proteins
4.4. Pheromone degrading enzymes and olfactory signal inactivation
4.4.1. Degradation of Y-lactones in scarab beetles
4.4.2. Degradation of the alkaloid pheromone from the pale brown chafer, P. diversa
5. Conclusions and perspective
6. Experimental procedures
6.1. Analysis of pheromones, their biosynthetic intermediates and degradation products by chromatographic methods
6.2. Protein purification and analysis
6.3. Ligand binding and enzymatic reactions with radiochemicals
6.4. Molecular cloning and protein expression
6.5. Spectroscopic analysis of protein structure
7. Literature cited
List of figures
Summary
Streszczenie